GENES CONTROLLING NEURTROPHIL DEVELOPMENT FROM STEM CELL

干细胞中控制嗜中性粒细胞发育的基因

• 批准号: 2149012
• 负责人: SCHICKWANN TSAI
• 金额: $ 13.23万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2149012
• 关键词: bone marrow transplantation; cell differentiation; genetic library; genetic regulation; hematopoietic stem cells; human tissue; laboratory mouse; leukopoiesis; neutrophil; nucleic acid hybridization; nucleic acid sequence; polymerase chain reaction; retinoate; tissue /cell culture; tissue mosaicism; transcription factor; transfection /expression vector

In this proposal, a novel murine lymphohematopoietic stem cell line MER and its derivative will be used to isolate genes controlling neutrophil commitment and differentiation. The MER cell line is dependent on stem cell factor (SCF; c-kit ligand) and has-both lymphoid and hematopoietic ("myeloid") potentials. From this stem cell line we established a neutrophilic derivative (MER-PRO) blocked at the promyelocyte stage. This block can be overcome with high concentrations of retinoic acid. This complementary pair of cell lines represent hematopoietic cells "frozen" reversibly at two developmental stages and are particularly suitable as a model system for studying stem cell commitment and lineage-restricted differentiation. The specific aims of this proposal are: (I) Isolation and characterization of neutrophil progenitor-specific genes, including those involved in lineage commitment. The technique of differential hybridization will be used to screen the cDNA library of MER-PRO (neutrophilic promyelocytes) using subtracted cDNA probes prepared from the MER-PRO and MER cell lines; (II) Isolation of genes activated or inactivated by retinoic acid during the terminal differentiation of neutrophilic promyelocytes. Either the cDNA library differential hybridization technique or the polymerase chain reaction-based "differential display" of mRNAs can be used. These genes are more downstream to those in Specific Aim I but are important in the later stages of neutrophil differentiation; (III) In vivo characterization of the lymphohematopoietic stem cell line MER and establishment of a human equivalent of the MER cell line. The capacity of the MER cell line to function as stem cells will be examined in irradiated syngeneic mice. Attempts will be made to create a similar cell line from human bone marrow. The MER cell line may be the first stem cell factor-dependent cell line with both lymphoid and hematopoietic potentials and should greatly facilitate stem cell research. The knowledge gained from this unique experimental system is likely to contribute to our understanding of the biology of human lymphohematopoietic stem cells. The long-term objective of this proposal is to identify molecules directly mediating the self-renewal, commitment and differentiation of stem cells and how these molecules are in turn modulated by hormones and extracellular matrix. It is hoped that this new knowledge will enable us to control stem cell behavior for therapeutic purposes such as gene therapy in the future.

在这项建议中，一种新的小鼠淋巴造血干细胞系MER 其衍生物将用于分离控制中性粒细胞的基因， 承诺和差异化。MER细胞系依赖于干细胞 细胞因子（SCF; c-kit配体），同时具有淋巴和造血功能 （“髓样”）电位。从这个干细胞系中，我们建立了一个 在早幼粒细胞阶段阻断嗜中性粒细胞衍生物（MER-PRO）。这 可以用高浓度的视黄酸来克服阻塞。这 一对互补的细胞系代表“冷冻”的造血细胞 在两个发育阶段是可逆的，并且特别适合作为 用于研究干细胞定型和谱系限制的模型系统 分化本建议的具体目标是：（一）隔离和 中性粒细胞祖细胞特异性基因的表征，包括那些 参与血统承诺。微分技术 杂交法筛选MER-PRO cDNA文库 （嗜酸性早幼粒细胞），使用从 MER-PRO和MER细胞系;（II）分离激活或 视黄酸灭活的终末分化过程中， 嗜酸性早幼粒细胞。cDNA文库差异 杂交技术或基于聚合酶链反应的 可以使用mRNA的“差异显示”。这些基因比 下游的具体目标，但在后者是重要的 中性粒细胞分化的阶段;（III） 淋巴造血干细胞系MER及人源性造血干细胞系的建立 等同于MER细胞系。MER细胞系的能力， 将在受辐射的同系小鼠中检查干细胞的功能。 将尝试从人骨中建立类似的细胞系 骨髓MER细胞系可以是第一个干细胞因子依赖性细胞 符合淋巴和造血潜能， 促进干细胞研究。从这个独特的 实验系统可能有助于我们了解 人类造血干细胞 这项提案的长期目标是直接识别分子 介导干细胞的自我更新、定型和分化 以及这些分子如何被激素调节， 细胞外基质希望这些新知识能使我们 控制干细胞行为用于治疗目的， 治疗在未来。